Certain steroid N-bis-(haloethyl)-carbamates

ABSTRACT

.Iadd.Antitumor active mono and poly N-Bis-(B-haloethyl)-carbamic acid esters of certain estrogenic steroid hormones, especially estradiol; such compounds wherein all hydroxyl groups of the steroid molecule are not thus esterified; and organic and inorganic acid esters of such latter compounds. .Iaddend.

The present invention relates to certain novel esters of certainestrogenic .[.androgenic and corticoid.]. steroid hormones, which estersshow an anti-tumour activity against Ehrlich's ascites tumours in mice.The acid moiety of these esters are acid residues ofN-bis-(β-haloethyl)-carbamic acids of the formula: ##STR1## wherein Xrepresents chlorine or bromine. The estrogenic .[.androgenic andcorticoid.]. steroid hormones that are used as starting materials forthe production of the said esters are simply termed "steroids" in thefollowing. They all contain one or more hydroxyl groups and may containfurther substituents such as .[.ether groups, halogen atoms, and.].alkyl groups. One or more of the hydroxyl groups may be previouslyesterified by introducing an acyl group of an unobjectionable acid, asrepresented by aliphatic, cycloaliphatic, and aromatic carboxylic acids,in particular mono- and dibasic carboxylic acids, as well as inorganicacids, such as phosphoric acids, phosphorous acids, sulfuric acids andthe like polybasic acids, leaving at least one hydroxyl group intact.

More particularly, the invention relates to mono- and polyesters ofestrogenic .[.androgenic and corticoid.]. steroid hormones selected fromthe group consisting of .[.estra-1,3,5 (10)-triene 17β-ol(3-desoxy-estradiol),.]. estra-1,3,5 (10)-triene-3,17β-diol(17β-estradiol), estra-1,3,5 (10)-triene-3,17α-diol (17α-estradiol),.[.estra-1,3,5 (10)-triene-3,17β-diol-16-one (16-keto-estradiol),.].estra-1,3,5 (10)-triene-3,16α, 17β-triol (estriol),17α-ethynyl-estra-1,3,5 (10)-triene-3,17β-diol(17α-ethynyl-17β-estradiol) .[.estra-1,3,5(10)-triene-3,17β-diol-3-methylether (17β-estradiol-3-methylether), andestra-1,3,5 (10)-triene-3,17β-diol-17-methylether(17β-estradiol-17-methylether), and androstane-3α, 17β-diol,androstane-3α-ol-17-one (androsterone), androstane-3β-ol-17-one(epiandrosterone), androst-5-ene-3β-ol-17-one (dehydroepiandrosterone),pregnane-3α-20α-diol (pregnanediol), etiocholane-3α -ol-17-one(etiocholanolone), pregn-5-ene-3β-ol-20-one pregnenolone),4-chloro-19-nor-androst-4-ene-17β-ol-3-one(4-chloro-19-nortestosterone), pregn-4-ene-17α,21-diol-3,11,20-trione(cortisone), pregn-4-ene-11β,17α,21-triol-3,20-dione (cortisol),pregn-4-ene-11β,21-diol-3,20-dione (corticosterone),pregn-4-ene-21-ol-3,11,20-trione (dehydrocorticosterone),pregn-4-ene-21-ol-3,20-dione (desoxycorticosterone),pregna-1,4-diene-17α,21-diol-3,11,20-trione (prednisone),pregna-1,4-diene-11β,17α,21-triol-3,20-dione (prednisolone),6α-methylpregna-1,4-diene-11β,17α,21-triol-3,20-dione(6α-methylprednisolone),9α-fluoro-pregn-4-ene-11β,17α,21-triol-3,20-dione (9α-fluorocortisol),6α-9α-difluoro-pregn-1,4-diene-11β,16α,17α,21tirol-3,20-dione(6α,9α-difluoro-prednisolone),16α-methyl-9α-fluoro-pregna-1,4-diene-11β,17α,21-triol-3,20-dione(16α-methyl-9α-fluoro-prednisolone).]. and the above-mentioned partialesters of these.

The esters of the invention are useful against Ehrlich's ascites tumorin mice, where a certain hormonal effect is desirable. This hormonaleffect, being inherent in the steroid moiety, can be regulated byintroducing at a remaining hydroxyl group in the steroid part of thenovel esters, an acid residue of the kind which is well known in the artfor regulating, modifying or protracting the effect of steroid hormones.The acids used for this purpose are the same as the unobjectionableorganic acids named above, and preferably such of not more than 20carbon atoms in the molecule.

The most promising compounds containing the estradiol nucleus areestradiol-3-N-bis-(β-chloroethyl)-carbamate,estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-phosphate,estradiol-3-N-bis-(β-chloroethyl)-carbamate 17-propionate,estradiol-3-phosphate-17-N-bis-(β-chloroethyl)-carbamate,estradiol-3,17-bis-[N-bis-(β-chloroethyl)]-carbamate,estradiol-17-N-bis-(β-chloroethyl)-carbamate,estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-acetate,estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-β(p-propoxyphenyl)-propionate,estradiol-3-acetate-17-N-bis-(β-chloroethyl)-carbamate,estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-trimethylacetate,estradiol-17-N-bis-(β-bromoethyl)-carbamate,estradiol-3-acetate-17-N-bis-(β-bromoethyl)-carbamate,estradiol-[3-N-bis-(β-chloroethyl)-17N-bis-(β-bromoethyl)]-dicarbamate.

The result of the three estradiol compounds first mentioned above astried against Ehrlich's ascites tumour is shown in the following Example.[.21..]. .Iadd.15..Iaddend.

Another object of the invention is to provide a process of preparing theabove-mentioned esters. When referring below to "esterificationaccording to the invention" what is meant is the introduction of acidresidues of Compound I into the steroid nucleus. The said esterificationcan be performed at one or several positions in the steroid nucleus, asdesired.

In a preferred embodiment of the process of the invention using asstarting materials:

(a) An estrogenic .[.androgenic or corticoid.]. steroid molecule asdefined hereinbefore with or without ester or .[.ether groups or.].other substituents at the nucleus, such as .[.halogen atoms or.]. alkylgroups, and containing a free hydroxyl group, and

(b) An N-bis (β-haloethyl)-amine of the general formula: ##STR2##wherein X is chlorine or bromine, one of the starting materials istreated with COY₂, wherein Y is chlorine or bromine, whereupon theresulting haloformyl derivative is reacted with the other startingmaterial, the resulting steroid ester being isolated and/or, if desired,being subsequently esterified with a compound selected from functionallyreactive derivatives of aliphatic, cycloaliphatic and aromaticcarboxylic acids, in particular mono- and dibasic carboxylic acids, aswell as inorganic acids, such as phosphoric acids, phosphorous acid,sulfuric acid or the like inorganic polybasic acids to introduce atleast one further ester group, the resulting ester being then isolated.

In the case of starting with an amine of Formula II, the preferred amineis the one, in which X represents chlorine.

The subsequent esterification process to introduce the acyl group of apolybasic inorganic acid is of particular interest, because it willgenerally result in products which may be made to dissolve in water toform aqueous solutions with a pH at or about the neutral point.

The esters of the invention may be administered, conveniently by theusual routes used for the administration of steroids, for example in theform of tablets, capsules or in the form of the usual sterile solutionsor suspensions for injection. The results upon administration have sofar been very promising.

The esters of the invention may be employed in aqueous suspension orsolution, or in oil solution or suspension, as well as dispersed inphysiological saline, but various preparations can also beadvantageously compounded which contain the active substance along withliquid or solid diluents. Solid preparations for extemporaneous dilutionas well as for direct usage (e.g., as implantates) may be formulated tocontain various buffering agents as well as local anesthetics and othermedicinal agents such as antibiotics, hypnotics, analgesics, etc., andinorganic salts to afford desirable pharmacological properties to thecomposition.

Doses of the order of 10 to 1000 mg./kg. daily and especially 50-400mg./kg. daily of the esters of the present invention are highlyeffective in inhibiting tumours in animals. Since the esters of theinvention are stable and widely compatible, they may be administered insolution or suspension in a variety of pharmacologically acceptablevehicles, including water, propylene glycol, diethyl, carbonate,glycerol, and oils such as peanut oil or sesame oil. Suitableformulations for compositions are as follows:

    ______________________________________                                        Tablet (implantate):                                                          Estradiol - 3 - N-bis-(β-chloroethyl)-carbamate,                         mg.                          100                                              Lactose, mg.                 100                                              Polyvinylpyrrolidone, mg.    4                                                Oil solution for injection:                                                   Estradiol - 3 - N-bis-(β-chloroethyl)-carbamate-                         17-propionate, mg.           250                                              Benzyl benzoate, mg.         15                                               Peanut oil to make, ml.      5                                                Aqueous suspension for injection:                                             Estradiol - 3 - N-bis-(β-chloroethyl)-carbamate,                         mg.                          250                                              Polyoxyethylene sorbitan monostearate (Tween                                  80, Atlas), mg.              20                                               Benzyl alcohol, mg.          45                                               Carboxymethyl-cellulose, mg. 15                                               Distilled water to make, ml. 5                                                Water solution for injection (can be lyophilized):                            Estradiol - 3 - N -bis-(β-chloroethyl)-carbamate-                        17-phosphate, mg.            250                                              Sodium hydroxide to make pH  7.0                                              Distilled water to make, ml. 10                                               ______________________________________                                    

Any other pharmaceutical adjuvants may be used provided that they arecompatible with the active ingredient.

In addition to the conventional intramuscular, subcutaneous,intravenous, and intraperitoneal administration routes, these compoundsmay also be employed in conjunction with perfusion procedures, whereinthe tumour site is isolated from the main circulatory system fortreatment.

As stated hereinbefore, the esterification according to the presentinvention may be accomplished using as starting materials a selectedsteroid, as hereinbefore defined, and a N-bis-(β-haloethyl)-amine,having the general formula: ##STR3## wherein X is as defined above. Oneof the starting materials is treated with COY₂, wherein Y representschlorine or bromine, whereupon the resulting haloformyl derivative isreacted with the other starting material and the resulting steroid ester(IV) is isolated (see Charts I and II below), if desired afteresterification with a functionally reactive derivative of apharmacologically unobjectionable acid as defined above. ##STR4##

The order of the addition of the reactants may influence the order atwhich introduction of the ester groups takes place at differentpositions in the steroid molecule. Thus using the order of additionshown in Chart I will favour the formation of an ester group at aphenolic hydroxyl group (such as the 3-OH group in estradiol) and at ahydroxyl group in 21-position in the steroid nucleus. By reversing theorder of addition as in Chart II, the formation of an ester group at ahydroxyl group in 3α,3β and 17 positions in the steroid nucleus isfavoured.

Some of the intermediate chloroformyl compounds (V) are novel. Theirmelting points (M.P.) and optical rotations ([α]_(D)) are given below:

    __________________________________________________________________________        Compound              M.P. ° C.                                                                     .[.[α]p°.].                                                           [α].sub.D°                 __________________________________________________________________________    17-chloroformate of estradiol-17β                                                                  123-124                                                                              +71.2                                        17-chloroformate of estradiol-17β-3-acetate                                                        112.5-114                                                                            +64.1                                        .[.3,17-dichloroformate of androstane-3α,17β                                                 147-147.5                                                                            +32.6.].                                     17 chloroformate of estradiol-17β-3-N-bis-(β-                       (chloroethyl)-carbamate   110-111.5                                                                            +44.8-.[.3-chloroformate of                                                   androstan-3α-ol-17-one                                                       119.5-120.5 +84.3.].                    .[.3-chloroformate of androst-5-ene-3β-ol-17-one                                                   126-127                                                                              +12.5.].                                     __________________________________________________________________________

It is convenient to carry out the esterification of the invention in thepresence of an acid-binding agent. As examples of such can be mentionedtertiary amines, such as anhydrous triethylamine, pyridine, quinolineand dimethylaniline, but also an excess of Compound II can serve asacid-binding agent.

It is also convenient to carry out the present esterification in thepresence of a solvent. Where the above-mentioned tertiary amines arepresent in excess, they may act as solvents; but otherwise an inertsolvent, such as anhydrous chloroform, benzene, toluene, xylene,tetrahydrofurane and dioxane, has been found very convenient. Otherinert solvents, such as high-boiling ethers, are also satisfactory.

The temperature is preferably kept at from about -20° C. up to about theboiling point of the mixture.

As functionally reactive derivatives of the pharmacologicallyunobjectionable acids mentioned above are preferably used acidanhydrides or acid chlorides, usually in an anhydrous, acid-bindingsolvent, such as triethylamine, pyridine, quinoline, dimethylaniline orthe like, at a temperature of preferably from -20° C. to about theboiling point of the mixture.

The following examples are given by way of illustration only and are notto be construed as limiting.

EXAMPLE 1 Estradiol-3-N-bis(β-chloroethyl)-carbamate

A solution in dry benzene of 82 g. of bis-(β-chloroethyl)-amine freshlyliberated from its hydrochloride is added gradually to a solution of 36g. of carbonyl chloride in benzene at a temperature below 10° C. Themixture is mechanically stirred for three hours, the precipitate ofbis-(β-chloroethyl)-amine hydrochloride is removed by filtration, andthe benzene is distilled off on a water bath. The residue is distilledin vacuo, and the N-chloroformyl-bis-(β-chloroethyl)-amine is obtainedas a pale yellow oil with a B.P. of 114-116° C./1 mm. Hg.

To a solution of 16.35 g. of estradiol in 75 ml. of dry pyridine, 21.00g. of the above-mentioned chloroformyl-bis-(β-chloroethyl)-amine areadded while stirring and cooling with ice-water.

The reaction mixture is allowed to stand at room temperature for 60-70hours under the exclusion of air humidity. Then the excess of thechloroformyl compound is hydrolyzed with crushed ice. Ethyl acetate isadded and, after shaking, the ethyl acetate solution is separated andwashed with water, dried over sodium sulphate and evaporated in vacuo todryness.

The residue is the 3-N-bis(β-chloroethyl)-carbamate of estradiol. Thecompound melts at 101-103° C. after recrystallization from isopropylether+hexane (1:1).

    [α].sub.D.sup.20.sup.°   C. =+ 44.8° (c.=1.0 in dioxane)

EXAMPLE 2 Estradiol-3-N-bis(β-chloroethyl)-carbamate-17-acetate

Analogously to Example 1,estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-acetate is obtained from6.28 g. of estradiol-17-acetate and 6.03 g. ofN-chloroformyl-bis-(β-chloroethyl)-amine. This compound melts at101-102° C. after recrystallization from methanol.

    [α].sub.D.sup.20.sup.°   C. =+ 23.8° (c.=1.0 in dioxane)

EXAMPLE 3Estradiol-3-N-bis(β-chloroethyl)-carbamate-17-β-(para-propoxyphenyl)-propionate

Analogously to Example 1,estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-β-(para-propoxyphenyl)-propionateis obtained from 4.63 g. ofestradiol-17-β-para-propoxyphenyl)-propionate and 3.08 g.N-chloroformyl-bis-(β-chloroethyl)-amine as an oil at room temperature.

    [α].sub.D.sup.20.sup.°   C. + 31.2° (c.=1.0 in dioxane)

.[.EXAMPLE 4 Cortisone-21-N-bis(β-chloroethyl)-carbamate

Analogously to Example 1, cortisone-21-N-bis-(β-chloroethyl)-carbamateis obtained from 1.80 g. of cortisone and 1.03 g. ofN-chloroformyl-bis-(β-chloroethyl)-amine. This compound melts at172.5-173.5° C. after recrystallization from methanol.

    [α].sub.D.sup.20.sup.°   C. =+ 179.2° (c.=1.0 in dioxane)

EXAMPLE 5 Prednisone-21-N-bis-(β-chloroethyl)-carbamate

Analogously to Example 1, prednisone-21-N-bis-(β-chloroethyl)-carbamateis obtained from 5.37 g. of prednisone and 3.06 g. ofN-chloroformyl-bis-(β-chloroethyl)-amine. This compound melts at180-181° C. after recrystallization from methanol.

    [α].sub.D.sup.20.sup.°  C. =+ 162.5° (c.=1.0 in dioxane)

EXAMPLE 6 Prednisolone-21-N-bis-(β-chloroethyl)-carbamate

Analogously to Example 1,prednisolone-21-N-bis-(β-chloroethyl)-carbamate is obtained from 5.40 g.of prednisolone and 3.06 g. of N-chloroformyl-bis-(β-chloroethyl)-amine.This compound melts at 179-180° C. after recrystallization from hexane.

    [α].sub.D.sup.20.sup.° C.=+109.7° (c.=1.0 in dioxane).].

EXAMPLE .[.7.]. .Iadd.4 .Iaddend.Androsterone-3-N-bis-(β-chloroethyl)-carbamate

To a solution of 7.05 g. of 17-chloroformic acid ester of androsteronein 85 ml. of dry chloroform is added a freshly produced solution of 6.25g. of bis-(β-chloroethyl)-amine in 80 ml. of dry chloroform (producedfrom 7.85 g. of the hydrochloride of bis-(β-chloroethyl)-amine) whilestirring and cooling with ice-water.

The reaction mixture must stand at room temperature for at least 6 hoursunder the exclusion of air humidity. The formed hydrochloride ofbis-(β-chloroethyl)-amine is then filtered off, whereupon the chloroformsolution is evaporated in vacuo to dryness.

The residue is the 3α-N-bis-(β-chloroethyl)-carbamate of androsterone.This compound melts at 109-110.5° C. after recrystallization fromhexane.

    [α].sub.D.sup.20.sup.° C.=+57.4° (c.=1.0 in dioxane)

.[.EXAMPLE 8 Dehydroepiandrosterone-3-N-bis-(β-chloroethyl)-carbamate

The above compound is obtained analogously to Example 7 from 3.51 g. of3β-1-chloroformic acid ester of dehydroepiandrosterone and 3.12 g. ofbis-(β-chloroethyl)-amine (produced from 3.93 g. of the hydrochloride).The compound melts at 129-130.5° C. after recrystallization from aqueousacetone.

    [α].sub.D.sup.20.sup.° C.=+18.9° (c.=1.0 in dioxane).].

EXAMPLE .[.9.]. .Iadd.5 .Iaddend.Estradiol-3-acetate-17-N-bis-(β-chloroethyl)-carbamate

Analogously to Example .[. 7.]. .Iadd.4.Iaddend., 17-N-bis-(β-chloroethyl)-carbamate of estradiol-3-acetate is obtained from 3.76 g.of 17-chloroformic acid ester of estradiol-3-acetate and 3.12 g. ofbis-(β-chloroethyl)-amine (produced from 3.93 g. of the hydrochloride).The compound melts at 98-99° C. after recrystallization from hexane.

    [α].sub.D.sup.20.sup.° C.=+55.8° (c.=1.0 in dioxane)

EXAMPLE .[.10.]. .Iadd.6 .Iaddend.Estradiol-17β-N-bis-(β-chloroethyl)-carbamate

To a solution of 0.5 g. potassium carbonate in 100 ml. aqueous methanolare added 1.44 g.estradiol-3-acetate-17β-N-bis-(β-chloroethyl)-carbamate (producedaccording to Example .[.9.]. .Iadd.5.Iaddend.) in a mixture of 65 ml.ethanol and 15 ml. acetone while stirring.

The reaction mixture is allowed to stand while stirring at a temperatureof about 20° C. for half an hour and poured into 200 ml. of cold 0.1 Nhydrochloric acid. After an hour, ethyl acetate is added and, aftershaking the ethyl acetate solution is separated and washed with water,dried over sodium sulphate and evaporated in vacuo to dryness.

The residue is the 17β-N-bis-(β-chloroethyl)-carbamate of estradiol. Thecompound melts at 138-139° C. after recrystallization from aqueousmethanol.

    [α].sub.D.sup.20.sup.° C.=+59.8° (c.=1.0 in dioxane)

.[.EXAMPLE 11 Androstane-3α,17β-bis-[N-bis-(β-chloroethyl)]-carbamate

This compound is obtained analogously to Example 7 from 0.83 g. of3α,17β-bis-chloroformic acid ester of androstane-3α,17β-diol and 1.25 g.of bis-(β-chloroethyl)-amine (produced from 1.57 g. of thehydrochloride). The compound melts at 87-88° C. after recrystallizationfrom isopropyl ether.

    [α].sub.D.sup.20.sup.° C.=+31.9° (c.=1.0 in dioxane).].

EXAMPLE .[.12.]. .Iadd.7.Iaddend.Estradiol-3,17-bis-[N-bis-(β-chloroethyl)]-carbamate

This compound is obtained analogously to Example .[.7.]. .Iadd.4.Iaddend. from 2.01 g. of 17β-chloroformic acid ester ofestradiol-3-N-bis(β-chloroethyl)-carbamate and 1.25 g. ofbis-(β-chloroethyl)-amine (produced from 1.57 g. of the hydrochloride).

The compound melts at 94.5-96.5° C. after recrystallization fromisopropyl ether.

    [α].sub.D.sup. 20.sup.° C.=+44.1° (c.=1.0 in dioxane)

EXAMPLE .[.13.]. .Iadd.8 .Iaddend.Estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-propionate

To a solution of 2.2 g. of 3-N-bis-(β-chloroethyl)-carbamate ofestradiol (produced according to Example 1) in 25 ml. of dry pyridine,6.5 g. of propionic acid anhydride are added while stirring and coolingwith ice-water. The reaction mixture is heated on a steam bath for 11/2hours under the exclusion of air humidity. After cooling, the surplus ofpropionic acid anhydride is hydrolyzed with crushed ice.

A mixture of equal parts of ether and ethyl acetate is added and, aftershaking, the organic phase is separated, washed with water and driedover sodium sulphate, whereupon it is evaporated in vacuo to dryness.The residue is 3-N-bis(β-chloroethyl)-carbamate-17-propionate ofestradiol. This compound melts at 70-72° C. after recrystallization fromaqueous methanol.

    [α].sub.D.sup.20.sup.° C.=+34.1° (c.=1.0 in dioxane)

EXAMPLE .[.14.]. .Iadd.9 .Iaddend.Estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-trimethyl-acetate

Analogously to Example .[.13.]. .Iadd.8.Iaddend., 3-N-bis-(β-chloroethyl)-carbamate-17-trimethylacetate of estradiol is obtainedfrom 2.2 g. of 3-N-bis-(β-chloroethyl)-carbamate of estradiol (producedaccording to example 1) and 6 ml. of pivalic acid chloride. The compoundmelts at 90-92° C. after recrystallization from aqueous methanol.

    [α].sub.D.sup.20.sup.° C.=+33.5° (c.=1.0 in dioxane)

EXAMPLE .[.15.]. .Iadd.10 .Iaddend.Estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-phosphate

To a solution of 2.3 ml. of phosphorus oxychloride in 50 ml. of drypyridine is added a solution of 2.2 g. of3-N-bis-(β-chloroethyl)-carbamate of estradiol while stirring and at atemperature of about -10° C. The reaction mixture is allowed to stand atabout 0° C. for one and a half hours, whereupon it is hydrolyzed bypouring it into ice water. The main part of the pyridine is evaporatedin vacuo, whereupon the residue is poured into 100 ml. of cold 3.5 Nhydrochloric acid with stirring. The precipitate thus obtained isisolated and washed with 0.1 N hydrochloric acid and water.

The compound, which consists of the 17-phosphate ofestradiol-3-N-bis(β-chloroethyl)-carbamate, melts under decomposition atabout 155° C. It is soluble in an aqueous solution of alkali.

    [α].sub.D.sup.20.sup.°  C. =+ 30.0° (c.=1.0 in dioxane)

EXAMPLE .[.16.]. .Iadd.11 .Iaddend.Estradiol-3-phosphate-17β-N-bis-(β-chloroethyl)-carbamate

Analogously to Example .[.15.]. .Iadd.10.Iaddend.,estradiol-3-phosphate-17β -N-bis-(β-chloroethyl)-carbamate is obtainedfrom 2.6 g. of 17β-N-bis-(β-chloroethyl)-carbamate of estradiol(produced according to Example .[.10.]. .Iadd.9 .Iaddend.and 2.75 ml. ofphosphorus oxychloride.

The compound melts under decomposition at about 125° C. It is soluble inan aqueous solution of alkali.

    [α].sub.D.sup.20.sup.°  C. =+ 49.9° (c=1.0 in chloroform)

.[.EXAMPLE 17 Androsterone-3α-N-bis-(β-bromoethyl)-carbamate

The above compound is obtained analogously to Example 7, from 1.00 g. ofandrosterone-3α-chloroformic acid ester and 1.43 g ofbis-(β-bromoethyl)-amine (produced from 1.94 g. of the hydrobromide).

The compound melts at 87.5-89° C. after recrystallization from isopropylether.

    [α].sub.D.sup.20.sup.°  C. =+ 56.7° (c.=1.0 in dioxane).].

EXAMPLE .[.18.]. .Iadd.12 .Iaddend.Estradiol-3-acetate-17-N-bis-(β-bromoethyl)-carbamate

Analogously to Example .[.7.]. .Iadd.4.Iaddend., the above compound isobtained from 1.25 g. of estradiol-3-acetate-17β-chloroformic acid esterand 1.68 g. of bis-(β-bromoethyl)-amine (obtained from 2.27 g. of thehydrobromide).

The compound melts at 95-96.5° C. after recrystallization from hexane.

    [α].sub.D.sup.20.sup.°  C. =+ 51.7° (c.=1.0 in dioxane)

EXAMPLE .[.19.]. .Iadd.13 .Iaddend.Estradiol-17-N-bis-(β-bromoethyl)-carbamate

Analogously to Example .[.7.]. .Iadd.4.Iaddend., the above compound isobtained from 1.75 g. of estradiol-17-β-chloroformic acid ester and 2.66g. of bis-(β-bromoethyl)-amine (obtained from 3.60 g. of thehydrobromide).

The compound melts at 149.5-151.5° C. after recrystallization fromisopropyl ether + hexane (1:1),

    [α].sub.D.sup.20.sup.°  C. =+ 66.3° (c.=1.0 in dioxane)

EXAMPLE .[.20.]. .Iadd.14 .Iaddend.Estradiol-3-N-bis-(β-chloroethyl)-17-N-bis-(β-bromoethyl)-dicarbamate

Analogously to Example .[.7.]. .Iadd.4.Iaddend., the above compound isobtained from 2.00 g. of 17-chloroformic acid ester ofestradiol-3-N-bis-(β-chloroethyl)-carbamate and 2.02 g. ofbis-(β-bromoethyl)-amine (obtained from 2.73 g. of the hydrobromide).

The compound melts at 74-78° C. after recrystallization from aqueousisopropyl alcohol.

    [α].sub.D.sup.20.sup.°  C. =+ 25.2° (c.=1.0 in dioxane)

EXAMPLE .[.21.]. .Iadd.15 .Iaddend. Animal experiment

The esters of the invention were tested for effectiveness in inhibitingthe growth of Ehrlich's ascites tumour according to the procedure laiddown in Cancer Chemother. Rep. (1959) 42-62. Administration was by theintraperitoneal route in each case. Some results obtained are given inTable 1.

                                      TABLE 1                                     __________________________________________________________________________                                  Tumour                                                                        Weight                                                              Dose ×                                                                       Leth-                                                                              treated/                                                            days,                                                                              ality,                                                                             control,                                            Compound        mg./kg.                                                                            percent                                                                            percent                                         __________________________________________________________________________    Estradiol-3-N-bis-(β-chloroethyl)-                                       carbamate           5 × 155                                                                      0    42                                              "                   5 × 412                                                                      0    10                                              Estradiol-3-N-bis-(β-chloroethyl)-                                       carbamate-17β-phosphate                                                                      5 × 66                                                                       0    42                                              "                   5 × 135                                                                      0    10                                              Estradiol-3-N-bis-(β-chloroethyl)-                                       carbamate-17β-propionate                                                                     5 × 400                                                                      0    30                                              __________________________________________________________________________

In Table 2 the estrogenic effect ofestradiol-3-N-bis-(β-chloroethyl)-carbamate is compared with that ofestradiol-3-benzoate using the duration (in days) of cornified vaginalsmears in spayed mice as an index of prolonged estrogenic activity. Eachanimal is injected with a single dose in oil. Ten animals are used ineach group.

                  TABLE 2                                                         ______________________________________                                                                 Dose    Dur-                                                                  in μg.                                                                             ation                                                                 steroid in                                           Compound                 equiv.  days                                         ______________________________________                                        Estrodiol-3-benzoate     8       8                                             "                       24      15                                            "                       72      18                                           Estradiol-3-N-bis(2-chloroethyl)-carbamate                                                             8       8                                             "                       24      15                                            "                       72      17                                           ______________________________________                                    

In Table 3 the estrogenic effect ofestradiol-3-N-bis-(2-chloroethyl)-carbamate-17-phosphate is comparedwith that of estradiol-17-phosphate using the weight of the uterus inspayed mice as an index of estrogenic activity four days after a singleinjection of a water solution of the sodium salts of the compounds. Tenanimals are used in each group.

                  TABLE 3                                                         ______________________________________                                                              Dose in   Uterus                                                              μg. steroid                                                                          weight                                            Compound          equiv.    in mg.                                        ______________________________________                                        Estradiol-17-phosphate                                                                              2         10                                                "                 10        13                                            Estradiol-3-N-bis-(β-chlorethyl)                                         carbamate-17-phosphate                                                                              2         19                                             "                    10        41                                            ______________________________________                                    

The high order of anti-tumour activity against Ehrlich's ascites tumourof the compounds of the present invention and compositions thereof istherefore evidenced by tests in lower mammals.

What we claim is:
 1. Mono- and polyesters of estrogenic steroid hormonesselected from the group consisting of.[.estra-1,3,5(10)-triene-17β-ol,.]. estra-1,3,5(10)-triene-3,17β-diol,estra-1,3,5(10)-triene-3,17α-diol,.[.estra-1,3,5(10)-triene-3,17β-diol-16-one,.].estra-1,3,5(10)-triene-3,16α,17β -triol,17α-ethynyl-estra-1,3,5(10)-triene-3,17β-diol, .[.estra-1,3,5(10)-triene-3,17β-diol-3-methyl-ether, and estra-1,3,5(10)-triene-3,17β-diol-17-methylether,.]. withN-bis-(β-haloethyl)-carbamic acid of the formula: ##STR5## wherein X isselected from the group consisting of chlorine and bromine, one or moreremaining hydroxyl groups when present in the steroid nucleus beingselected from the group consisting of free hydroxyl groups and hydroxylgroups esterified with an acid selected from the group consisting ofaliphatic, cycloaliphatic and aromatic carboxylic acids, and inorganicpolybasic acids.
 2. The esters of claim 1, in which the sole acid moietyis N-bis-(β-haloethyl)-carbamic acid (compound I).
 3. The esters ofclaim 1, in which the hormone is estradiol. 4.Estradiol-3-N-bis-(β-chloroethyl)-carbamate. 5.Estradiol-3,17-bis-N-bis-(β-chloroethyl)-carbamate. 6.Estradiol-17-phosphate-3-N-bis-(β-chloroethyl)-carbamate. 7.Estradiol-3-phosphate-17-N-bis-(β-chloroethyl)-carbamate. 8.Estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-propionate. .[.9. Mono-and polyesters of androgenic steroid hormones selected from the groupconsisting of androstane-3α,17β-diol, androstane-3α-ol-17-one,androstane-3β-ol-17-one, androst-5-ene-3β-ol-17-one, andetiocholane-3α-ol-17-one, with N-bis-(β-haloethyl)-carbamic acid of theformula: ##STR6## wherein X is selected from the group consisting ofchlorine and bromine, one or more remaining hydroxyl groups when presentin the steroid nucleus being selected from the group consisting of freehydroxyl groups and hydroxyl groups esterified with an acid selectedfrom the group consisting of aliphatic, cycloaliphatic and aromaticcarboxylic acids, and inorganic polybasic acids..]. .[.10. Mono- andpolyesters of corticoid steroid hormones selected from the groupconsisting of pregnane-3α,20α-diol, pregn-5-ene-3β-ol-20-one,pregn-4-ene-17α, 21-diol-3,11-20-trione,pregn-4-ene-11β,17α,21-triol-3,20-dione, pregn-4-ene-11β,21-diol-3,20-dione, pregn-4-ene-21-ol-3,11-20-trione,pregn-4-ene-21-ol-3,20-dione, pregn-1,4-diene-17α21-diol-3,11,20-trione,pregna-1,4-diene-11β,17α,21-triol-3,20-dione,6α-methyl-pregna-1,4-diene-11β,17α,21-triol-3,20-dione,9α-fluoro-pregn-4-ene-11β,17α,21-triol-3,20-dione,6α9α-difluoro-pregn-1,4-diene-11β,16α,17α,21-tetrol-3,20-dione and16α-methyl-9α-fluoro-pregna-1,4-diene-11β,17α,21-triol-3,20-dione withN-bis(β-haloethyl)-carbamic acid of the formula: ##STR7## wherein X isselected from the group consisting of chlorine and bromine, one or moreremaining hydroxyl groups when present in the steroid nucleus beingselected from the group consisting of free hydroxyl groups and hydroxylgroups esterified with an acid selected from the group consisting ofaliphatic, cycloaliphatic and aromatic carboxylic acids, and inorganicpolybasic acids..]. 11.Estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-lower-alkanoate. .Estradiol-17β-N-bis-(β-chloroethyl)-carbamate. 13.Estradiol-17-N-bis-(β-bromoethyl)-carbamate. 14.Estradiol-3-acetate-17-N-bis-(β-chloroethyl)-carbamate. 15.Estradiol-3-lower-alkanoate-17-N-bis-(β-chloroethyl)-carbamate. .Estradiol-3-acetate-17-N-bis-(β-bromoethyl)-carbamate. 17.Estradiol-3-lower-alkanoate-17-N-bis-(β-bromoethyl)-carbamate.Estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-trimethyl-acetate. 9.Estradiol-3-N-bis-(β-chloroethyl)-carbamate-17-acetate.